Several classes of active compounds have already been described for the treatment of obesity and disorders of lipid metabolism:
polymeric adsorbers, such as, for example, cholestyramine PA1 benzothiazepines (WO 93/16055) PA1 bile acid dimers and conjugates EP 0 489 423) PA1 4-amino-2-ureidopyrimidine-5-carboxamides (EP 0 557 879) PA1 R.sup.3 to R.sup.8 independently of one another are hydrogen, fluorine, chlorine, bromine, iodine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(R.sup.9)R.sup.10, CHO, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12, it being possible in the alkyl radicals for one or more hydrogen(s) to be replaced by fluorine; PA1 R.sup.9 to R.sup.12 independently of one another are hydrogen, (C.sub.1 -C.sub.8)-alkyl; PA1 X is CH, NH; PA1 Y is CH, NH; PA1 R.sup.1 is phenyl; PA1 R.sup.2 is phenyl; PA1 X is CH; PA1 Y is CH; PA1 R.sup.1 and R.sup.2 independently of one another are cycloalkyl having 3-8 ring carbon atoms, phenyl, naphthyl, thienyl, furyl, pyrimidyl, thiazolyl, imidazolyl, phthalimidyl, quinolyl, piperazinyl, tetrazolyl, triazolyl, oxazolyl or their thieno-, pyridino- or benzo-fused derivatives, it being possible for the cycloalkyl ring, aromatic ring or heteroaromatic ring to be mono- to tri-substituted by fluorine, chlorine, bromine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(R.sup.9)R.sup.10, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12, it being possible in the alkyl radicals for one or more hydrogen(s) to be replaced by fluorine; PA1 R.sup.3 to R.sup.8 independently of one another are hydrogen, fluorine, chlorine, bromine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(R.sup.9)R.sup.10, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12, it being possible in the alkyl radicals for one or more hydrogen(s) to be replaced by fluorine; PA1 R.sup.9 to R.sup.12 independently of one another are hydrogen, (C.sub.1 -C.sub.8)-alkyl; PA1 X is CH, NH; PA1 Y is CH, NH; PA1 R.sup.1 is phenyl; PA1 R.sup.2 is phenyl; PA1 X is CH; PA1 Y is CH; PA1 R.sup.1 is pyridyl, pyrimidyl, thienyl, thiazolyl, it being possible for the heteroaromatic ring to be mono- to trisubstituted by fluorine, chlorine, bromine, iodine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(R.sup.9)R.sup.10, CHO, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12 ; PA1 R.sup.2 is phenyl, it being possible for the aromatic ring to be mono- to trisubstituted by fluorine, chlorine, bromine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(.sup.9)R.sup.10, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12 ; PA1 R.sup.3 to R.sup.8 independently of one another are hydrogen, fluorine, chlorine, bromine, iodine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(.sup.9)R.sup.10, CHO, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12, it being possible in the alkyl radicals for one or more hydrogen(s) to be replaced by fluorine; PA1 R.sup.9 to R.sup.12 independently of one another are hydrogen, (C.sub.1 -C.sub.8)-alkyl; PA1 X is CH; PA1 Y is NH;
The invention is based on the object of making availiable compounds which display a therapeutically utilizable hypolipidemic action.
The invention therefore relates to propanolamine derivatives of the formula I, ##STR2## in which R.sup.1 and R.sup.2 independently of one another are cycloalkyl having 3-8 ring carbon atoms, phenyl, naphthyl, phenanthryl, pyridyl, thienyl, furyl, pyrimidyl, indolyl, thiazolyl, imidazolyl, coumarinyl, phthalimidyl, quinolyl, piperazinyl, tetrazolyl, triazolyl, oxazolyl or their thieno-, pyridino- or benzo-fused derivatives, it being possible for the cycloalkyl ring, aromatic ring or heteroaromatic ring to be mono- to trisubstituted by fluorine, chlorine, bromine, iodine, OH, CF.sub.3, --NO.sub.2, CN, (C.sub.1 -C.sub.8)-alkoxy, (C.sub.1 -C.sub.8)-alkyl, NH.sub.2, --NH--R.sup.9, --N(R.sup.9)R.sup.10, CHO, --COOH, --COOR.sup.11, --(C.dbd.O)--R.sup.12, (C.sub.1 -C.sub.6)-alkyl-OH, (C.sub.1 -C.sub.6)-alkyl(--OH)-phenyl, (C.sub.1 -C.sub.6)-alkyl-CF.sub.3, (C.sub.1 -C.sub.6)-alkyl-NO.sub.2, (C.sub.1 -C.sub.6)-alkyl-CN, (C.sub.1 -C.sub.6)-alkyl-NH.sub.2, (C.sub.1 -C.sub.6)-alkyl-NH--R.sup.9, (C.sub.1 -C.sub.6)-alkyl-N(R.sup.9)R.sup.10, (C.sub.1 -C.sub.6)-alkyl-CHO, (C.sub.1 -C.sub.6)-alkyl-COOH, (C.sub.1 -C.sub.6)-alkyl-COOR.sup.11, (C.sub.1 -C.sub.6)-alkyl-(C.dbd.O)--R.sup.12, --O--(C.sub.1 -C.sub.6)-alkyl-OH, --O--(C.sub.1 -C.sub.6)-alkyl-CF.sub.3, --O--(C.sub.1 -C.sub.6)-alkyl-NO.sub.2, --O--(C.sub.1 -C.sub.6)-alkyl-CN, --O--(C.sub.1 -C.sub.6)-alkyl-NH.sub.2, --O--(C.sub.1 -C.sub.6)-alkyl-NH--R.sup.9, --O--(C.sub.1 -C.sub.6)-alkyl-N(R.sup.9)R.sup.10, --O--(C.sub.1 -C.sub.6)-alkyl-CHO, --O--(C.sub.1 -C.sub.6)-alkyl-COOH, --O--(C.sub.1 -C.sub.6)-alkyl-COOR.sup.11, --O--(C.sub.1 -C.sub.6)-alkyl-(C.dbd.O)--R.sup.12, --N--SO.sub.3 H, --SO.sub.2 --CH.sub.3, --O--(C.sub.1 -C.sub.6)-alkyl-O--(C.sub.1 -C.sub.6)-alkylphenyl, it being possible in the alkyl radicals for one or more hydrogen(s) to be replaced by fluorine;
with the proviso that the radicals R.sup.1, R.sup.2, X and Y do not simultaneously have the following meaning:
and their physiologically tolerable acid addition salts.
Preferred compounds of formula I are those in which one or more radical(s) has or have the following meaning:
with the proviso that the radicals R.sup.1, R.sup.2, X and Y do not simultaneously have the following meaning:
and their physiologically tolerable acid addition salts.
Particularly preferred compounds of formula I are those in which one or more radical(s) has or have the following meaning:
and their physiologically tolerable acid addition salts.
Physiologically tolerable acid addition salts are understood as meaning readily water-soluble, soluble and less soluble compounds according to the definition in Deutschen Arzneibuch ("German Pharmacopeia", 9th Edition 1986, Official Edition, Deutscher Apotheker-Verlag Stuttgart), page 19. The hydrochlorides and sulfates of the compounds are preferred.
The invention relates both to isomer mixtures of formula I, and to the pure enantiomers of formula I.
The invention furthermore relates to processes for the preparation of propanolamine derivatives of formula I. ##STR3##
Process A for the preparation of the compounds of formula I comprises a) preparing imines substituted by R.sup.1 and R.sup.2 and which are unknown from the literature, R.sup.1 and R.sup.2 having the meaning indicated for formula I, following literature processes, from amines of type II and aldehydes of type III. To do this, for example, the amine II and the aldehyde III are reacted in undiluted form or in a suitable solvent such as ethanol, toluene or acetic acid with or without addition of an acid, e.g. p-toluenesulfonic acid, at temperatures of 20.degree.-150.degree. C.
Keto compounds of formula VII substituted by radicals R.sup.3 to R.sup.8, R.sup.3 to R.sup.8 having the meaning indicated for formula I, are prepared by processes known from the literature or following such processes. Thus, for example, picoline derivatives V are metalated with a suitable base, such as n-butyllithium, and reacted in tetrahydrofuran or another suitable solvent with the corresponding carboxylic acid derivatives VI, e.g. present as carboxylic acid dialkylamides or esters, at temperatures between -80.degree. and 20.degree. C.
Compounds of type VIII are obtained by reacting imines of type IV and ketones of type VII, in each case substituted by radicals R.sup.3 to R.sup.8, R.sup.3 to R.sup.8 having the meaning indicated for formula I. This reaction can be carried out, for example, by mixing of the two compounds in diluted form, without solvent, and subsequent heating, or in a suitable solvent such as ethanol, toluene, diglyme or tetradecane, at temperatures from 20.degree. C. to 150.degree. C. (c).
The keto compounds of type VIII are reduced to hydroxy compounds of type I in a suitable solvent, such as, for example, methanol, THF or THF/water using NaBH.sub.4 or another suitable reductant, at temperatures between -30.degree. and +40.degree. C., it being possible for compound I to be substituted by the radicals R.sup.3 to R.sup.8 and R.sup.3 to R.sup.8 having the meaning indicated for formula I (d).
The compounds of formula I are obtained by the above-described reduction as isomer mixtures. Different isomers can be separated from one another by fractional crystallization or by column chromatography. The pure enantiomers can be obtained from the racemates of the compounds of formula I by chromatography on chiral column material or by processes known from the literature using optically active auxiliary reagents, such as described in J. Org. Chem. 44, 4891 (1979). In addition, the compounds of formula I can be readily converted into their physiologically tolerable acid addition salts by known processes.
Process B for the preparation of the compounds of formula I comprises not preparing and isolating the imine compound IV as process A, but preparing the compounds of type VIII, substituted by the radicals R.sup.3 to R.sup.8, in a three-component reaction from ketones VII, amines II and aldehydes III. To do this, these three components are reacted in undiluted form or in a suitable solvent, such as ethanol, tetradecane or toluene, at temperatures from 20.degree. C. to 150.degree. C. (e). The compounds VIII are reduced, as described for process A, to the compounds of formula I (f), it being possible to employ the compounds VIII as purified ketones, but also as crude products from the reaction described above.
The present invention also relates to pharmaceutical preparations which, in addition to nontoxic, inert, pharmaceutically suitable excipients, contain one or more active compounds according to the invention or which consist of one or more active compounds according to the invention, and to processes for the production of these preparations.
Nontoxic, inert, pharmaceutically suitable excipients are pharmaceutically acceptable solid, semisolid or liquid diluents, fillers and formulation auxiliaries of any type, which after mixing with the active compound bring this into a form suitable for administration.
Suitable administration forms of the compounds according to the invention are, for example, tablets, coated tablets, capsules, pills, aqueous solutions, suspensions and emulsions, where appropriate sterile injectable solutions, nonaqueous emulsions, suspensions and solutions, sprays and also preparation forms having protracted release of active compound.
The therapeutically active compounds should be present in the above mentioned pharmaceutical preparations expediently in a concentration from approximately 0.1 to 99.0, preferably from 0.5 to 70.0, percent by weight of the total mixture.
The administration concentrations for solutions and also aerosols in the form of sprays are in general 0.1 to 20, preferably 0.5 to 5, percent by weight.
Apart from the active compounds according to the invention, the abovementioned pharmaceutical preparations can also contain other pharmaceutically active compounds.
The abovementioned pharmaceutical preparations are prepared in a customary manner by known methods, e.g. by mixing the active compound(s) with the excipient(s).
The active compounds or the pharmaceutical preparations can be administered orally, parenterally, intraperitoneally and/or rectally.
The compounds of the present invention and their salts, which are utilizable, for example, as hypolipidemics, can be used for the production of pharmaceutical preparations which contain an efficacious amount of the active substance together with excipients and which are suitable for enteral and parenteral administration. Tablets or capsules (gelatin capsules) are preferably used which contain the active compound together with diluents or excipients, e.g. lactose, dextrose, cane sugar, mannitol, sorbitol, cellulose, various types of starch and/or glycine, and lubricants such as silica, talc, stearic acid or its salts, such as magnesium or calcium stearate, and/or polyethylene glycol. Tablets also contain binders such as magnesium carbonate, magnesium aluminum silicate, starch, gelatine, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone and, if required, colorants, flavorings and sweeteners. Injectable solutions are preferably isotonic aqueous solutions or suspensions, which can be sterilized and can contain auxiliaries, such as preservatives, stabilizers, wetting agents and/or emulsifiers, solubilizers, salts for regulating the osmotic pressure and/or buffer substances. The pharmaceutical preparations according to the invention, which if desired can contain further pharmacologically active substances, are produced, for example, by means of conventional mixing, granulating and sugar-coating processes and contain 0.1% to 80%, preferably approximately 5% to approximately 65%, of the active compound.
Oral administration takes place in pharmaceutically customary preparations, for example in the form of tablets, coated tablets or capsules which contain, for example, per daily dose 5 to 1000 mg, preferably 20 to 200 mg, of the active compound as a mixture with a customary excipient and/or constituent, it being possible to give individual doses of 5 to 200 mg, preferably one to three times daily.
However, it may be necessary to depart from the doses mentioned, namely depending on the type and the body weight of the subject to be treated, the nature and severity of the disorder, the type of preparation and the administration of the medicament, and the time or interval within which administration takes place. Thus in some cases it may be adequate to manage with less than the abovementioned amount of active compound, while in other cases the abovementioned amount of active compound must be exceeded. The optimal dose and type of administration of the active compounds necessary in each case can be established by any person skilled in the art on account of his/her expert knowledge.
The compounds of formula I and their physiologically tolerable salts are ideal pharmaceuticals for the treatment of disorders of lipid metabolism, in particular of hyperlipidemia. The compounds of formula I are also suitable for affecting the serum cholesterol level and for the prevention and treatment of arteriosclerotic symptoms. The following findings confirm the pharmacological activity of the compounds according to the invention.
The biological testing of the compounds according to the invention was carried out by determination of the inhibition of the .sup.3 H!-taurocholate uptake in brush-border membrane vesicles of the ileum of rabbits. The inhibition test was carried out as follows:
1. Preparation of brush-border membrane vesicles from the ileum of rabbits
The preparation of brush-border membrane vesicles from the intestinal cells of the small intestine was carried out using the so-called Mg.sup.2+ precipitation method. Male New Zealand rabbits (2 to 2.5 kg body weight) were sacrificed by intravenous injection of 0.5 ml of T61.RTM., an aqueous solution of 2.5 mg of tetracaine HCl, 100 mg of embutramide and 25 mg of mebezonium iodide. The small intestine was removed and rinsed with ice-cold physiological saline solution. The terminal 7/10 of the small intestine (measured in the oral-rectal direction, i.e. the terminal ileum, which contains the active Na.sup.+ -dependent bile acid transport system) were used for the preparation of the brush-border membrane vesicles. The intestines were frozen in plastic bags under nitrogen at -80.degree. C. For the preparation of the membrane vesicles, the frozen intestines were thawed at 30.degree. C. in a water bath. The mucosa were scraped off and suspended in 60 ml of ice-cold 12 mM Tris/HCl buffer (pH 7.1)/300 mM mannitol, 5 mM EGTA/10 mg/L of phenylmethylsulfonyl fluoride/L mg/L of trypsin inhibitor from soybeans (32 U/mg)/0.5 mg/L trypsin inhibitor from bovine lung (193 U/mg)/5 mg/L of bacitracin. After diluting to 300 ml with ice-cold distilled water, homogenization was carried out with an Ultraturrax (18-rod, IKA Werk Staufen, Germany) for 3 minutes at 75% max. power with ice-cooling. After addition of 3 ml of 1M MgCl.sub.2 solution (final concentration 10 mM), the homogenizate was allowed to stand at 0.degree. C. for exactly 1 minute. By addition of Mg.sup.2+, the cell membranes aggregate and precipitate with the exception of the brush-border membranes. After centrifugation at 3000.times.g for 15 minutes (5000 rpm, SS-34 rotor), the precipitate was discarded and the supernatant which contains the brush-border membranes was centrifuged at 48000.times.g for 30 minutes (20000 rpm, SS-34 rotor). The supernatant was discarded and the precipitate was re-homogenized in 60 ml of 12 mM Tris/HCl buffer (pH 7.1)/60 mM mannitol, 5 mM EGTA using a Potter Elvejhem homogenizer (Braun Melsungen, 900 rpm, 10 strokes). After addition of 0.1 ml of 1M MgCl.sub.2 solution and an incubation time of 15 minutes at 0.degree. C., centrifugation at 3000.times.g was again carried out for 15 minutes. The supernatant was then centrifuged again at 48000.times.g for 30 minutes (20000 rpm, SS-34 rotor). The precipitate was taken up in 30 ml of 10 mM Tris/HEPES buffer (pH 7.4)/300 mM mannitol and homogeneously resuspended by 20 strokes in a Potter Elvejhem homogenizer at 1000 rpm. After centrifugation at 48000.times.g for 30 minutes (20000 rpm, SS-34 rotor), the prcipitate was taken up in 0.5 to 2 ml of Tris/HEPES buffer (pH 7.4)/280 mM mannitol (final concentration 20 mg/ml) and resuspended with the aid of a tuberculin syringe having a 27 gauge needle. The vesicles were either used for transport investigations immediately after preparation or stored in liquid nitrogen at -196.degree. C. in 4 mg aliquots.
2. Inhibition of the Na.sup.+ -dependent .sup.3 !-taurocholate absorption in brush-border membrane vescles of the ileum
The absorption of substrates in the brush-border membrane vesicles described above was determined by means of the so-called membrane filtration technique. 10 .mu.l of the vesicle suspension (100 .mu.g of protein) were pipetted as drops onto the wall of a polystyrene incubation tube (11.times.70 mm) which contained the incubation medium with the appropriate ligands (90 .mu.l). The incubation medium contained 0.75 .mu.l=0.75 .mu.Ci of .sup.3 H(G)!-taurocholate (specific activity: 2.1 Ci/mmol)/0.5 .mu.l of 10 mM taurocholate/8.75 .mu.l of sodium transport buffer (10 mM Tris/HEPES (pH 7.4)/100 mM mannitol/100 mM NaCl) (Na-T-B) or 8.75 .mu.l of potassium transport buffer (10 mM tris/HEPES (pH 7.4)/100 mM mannitol/100 mM KCl) (K-T-B) and 80 .mu.l of the inhibitor solution concerned, dissolved in Na-T-buffer or K-T-buffer depending on the experiment. The incubation medium was filtered through a polyvinylidene fluoride membrane filter (SYHV LO 4NS, 0.45 .mu.m, 4 mm diameter, Millipore, Eschborn, Germany). Transport measurement was started by mixing the vesicles with the incubation medium. The concentration of taurocholate in the incubation batch was 50 .mu.M. After the desired incubation time (customarily 1 minute), the transport was stopped by addition of 1 ml of ice-cold stop solution (10 mM tris/HEPES, (pH 7.4)/150 mM KCl). The resulting mixture was immediately filtered off under a vacuum of 25 to 35 mbar through a membrane filter of cellulose nitrate (ME 25, 0.45 .mu.m, 25 mm diameter, Schleicher & Schuell, Dassell, Germany). The filter was washed with 5 ml of ice-cold stop solution.
For measurement of the absorption of the radiolabeled taurocholate, the membrane filter was dissolved using 4 ml of the scintillator Quickszint 361 (Zinsser Analytik GmbH, Frankfurt, Germany) and the radioactivity was measured by liquid scintillation measurement in a TriCarb 2500 measuring apparatus (Canberra Packard GmbH, Frankfurt, Germany). The measured values were obtained as dpm (decompositions per minute) after calibration of the apparatus with the aid of standard samples and after correction for any chemiluminescence present.
The control values were in each case determined in Na-T-B and K-T-B. The difference between the absorption in Na-T-B and K-T-B gave the Na.sup.+ -dependent transport fraction. The IC.sub.50 Na.sup.+ was designated as that concentration of inhibitor at which the Na.sup.+ -dependent transport fraction was inhibited by 50%--relative to control.
The pharmacological data comprise a series of tests in which the interaction of the compounds according to the invention with the intestinal bile acid transport system was investigated in the terminal small intestine. The results are summarized in Table 1.
Table 1 shows measurements of the inhibition of the .sup.3 H!-taurocholate absorption in brush-border membrane vesicles of the ileum of rabbits. The quotients of the IC.sub.50 Na values of the reference substance and taurochenodeoxycholate (TCDC) and the respective test substance are indicated.
TABLE 1 ______________________________________ Compounds from Example ##STR4## ______________________________________ 6 0.10 10 0.36 32 0.29 36 0.22 61 0.20 70 0.27 72 0.28 83 0.22 86 0.24 101 0.23 ______________________________________
The following Examples serve to illustrate the invention in greater detail without restricting the latter to products and embodiments described in the Examples.